The novel method of obtaining cells’ electrorotation motion parameters through electrorotation test was proposed. Firstly, constructing the cell edge detection algorithm to locate edges, then, the ring window template was used to split every single cell from multicellular images by means of template matching scheme. Feature point tracking and cross-correlation tracking algorithms for cell rotation rate measurement were proposed, and detection performance were compared. The processing results of the electro-rotation video images shows, the accurate cell segmentation was realized through the ring window template matching, the rotation rate of cell could be measured effectively in both methods, under the same measurement accuracy, cross-correlation tracking method owns better effect than feature point tracking.